Aims To evaluate ATP bioluminescence‐based hygiene monitoring systems under conditions relevant to fish processing environments. Methods and Results The ATP bioluminescence of fish fractions that are potentially present after insufficient cleaning of fish processing environments was determined. Different fractions and interfering substances representing the stages from slaughtering to smoking were prepared and measured using two different commercial systems (SystemSURE Plus and Clean‐Trace). ATP bioluminescence was quenched by acidic liquid smoke and by sodium chloride even at concentrations as low as 0·9% NaCl. Large variations were observed between different types of trout homogenates: the ATP bioluminescence from raw belly fat homogenate was 100–1000 times lower than for trout blood. There were about a 1000‐fold lower ATP bioluminescence in raw compared to heat‐treated fractions from trout, with the exception of blood. The bioluminescence from Listeria monocytogenes was very low. Results from fish processing plants supported the laboratory findings. Conclusions The output from ATP‐monitoring instruments depends on the nature of fish soil present, as well as the presence of sodium chloride and low pH. This may lead to considerable under‐ or overestimation of the level of organic soil. Significance and Impact of the Study ATP bioluminescence instruments are widely used by the fish industry for monitoring hygiene. The monitoring method will only give valuable information about the hygiene if critical limits are set after a validation period, distinguishing between areas with different types of soil and between different hygiene zones.
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